Development and validation of protein microarray technology for simultaneous inflammatory mediator detection in human sera
Biomarkers, including cytokines, can help in the diagnosis, prognosis, and prediction of treatment response across a wide range of disease settings. Consequently, the recent emergence of protein microarray technology, which is able to quantify a range of inflammatory mediators in a large number of s...
| Main Authors: | , , , , , , , |
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| Format: | Article |
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Hindawi
2014
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| Online Access: | https://eprints.nottingham.ac.uk/40717/ |
| _version_ | 1848796123770126336 |
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| author | Selvarajah, Senthooran Negm, Ola H. Hamed, Mohamed R. Tubby, Carolyn Todd, Ian Tighe, Patrick J. Harrison, Tim Fairclough, Lucy C. |
| author_facet | Selvarajah, Senthooran Negm, Ola H. Hamed, Mohamed R. Tubby, Carolyn Todd, Ian Tighe, Patrick J. Harrison, Tim Fairclough, Lucy C. |
| author_sort | Selvarajah, Senthooran |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Biomarkers, including cytokines, can help in the diagnosis, prognosis, and prediction of treatment response across a wide range of disease settings. Consequently, the recent emergence of protein microarray technology, which is able to quantify a range of inflammatory mediators in a large number of samples simultaneously, has become highly desirable. However, the cost of commercial systems remains somewhat prohibitive. Here we show the development, validation, and implementation of an in-house microarray platform which enables the simultaneous quantitative analysis of multiple protein biomarkers. The accuracy and precision of the in-house microarray system were investigated according to the Food and Drug Administration (FDA) guidelines for pharmacokinetic assay validation. The assay fell within these limits for all but the very low-abundant cytokines, such as interleukin- (IL-) 10. Additionally, there were no significant differences between cytokine detection using our microarray system and the “gold standard” ELISA format. Crucially, future biomarker detection need not be limited to the 16 cytokines shown here but could be expanded as required. In conclusion, we detail a bespoke protein microarray system, utilizing well-validated ELISA reagents, that allows accurate, precise, and reproducible multiplexed biomarker quantification, comparable with commercial ELISA, and allowing customization beyond that of similar commercial microarrays. |
| first_indexed | 2025-11-14T19:42:59Z |
| format | Article |
| id | nottingham-40717 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:42:59Z |
| publishDate | 2014 |
| publisher | Hindawi |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-407172020-05-04T16:55:59Z https://eprints.nottingham.ac.uk/40717/ Development and validation of protein microarray technology for simultaneous inflammatory mediator detection in human sera Selvarajah, Senthooran Negm, Ola H. Hamed, Mohamed R. Tubby, Carolyn Todd, Ian Tighe, Patrick J. Harrison, Tim Fairclough, Lucy C. Biomarkers, including cytokines, can help in the diagnosis, prognosis, and prediction of treatment response across a wide range of disease settings. Consequently, the recent emergence of protein microarray technology, which is able to quantify a range of inflammatory mediators in a large number of samples simultaneously, has become highly desirable. However, the cost of commercial systems remains somewhat prohibitive. Here we show the development, validation, and implementation of an in-house microarray platform which enables the simultaneous quantitative analysis of multiple protein biomarkers. The accuracy and precision of the in-house microarray system were investigated according to the Food and Drug Administration (FDA) guidelines for pharmacokinetic assay validation. The assay fell within these limits for all but the very low-abundant cytokines, such as interleukin- (IL-) 10. Additionally, there were no significant differences between cytokine detection using our microarray system and the “gold standard” ELISA format. Crucially, future biomarker detection need not be limited to the 16 cytokines shown here but could be expanded as required. In conclusion, we detail a bespoke protein microarray system, utilizing well-validated ELISA reagents, that allows accurate, precise, and reproducible multiplexed biomarker quantification, comparable with commercial ELISA, and allowing customization beyond that of similar commercial microarrays. Hindawi 2014-10-14 Article PeerReviewed Selvarajah, Senthooran, Negm, Ola H., Hamed, Mohamed R., Tubby, Carolyn, Todd, Ian, Tighe, Patrick J., Harrison, Tim and Fairclough, Lucy C. (2014) Development and validation of protein microarray technology for simultaneous inflammatory mediator detection in human sera. Mediators of Inflammation, 2014 . 820304/1-820304/12. ISSN 1466-1861 https://www.hindawi.com/journals/mi/2014/820304/ doi:10.1155/2014/820304 doi:10.1155/2014/820304 |
| spellingShingle | Selvarajah, Senthooran Negm, Ola H. Hamed, Mohamed R. Tubby, Carolyn Todd, Ian Tighe, Patrick J. Harrison, Tim Fairclough, Lucy C. Development and validation of protein microarray technology for simultaneous inflammatory mediator detection in human sera |
| title | Development and validation of protein microarray technology for simultaneous inflammatory mediator detection in human sera |
| title_full | Development and validation of protein microarray technology for simultaneous inflammatory mediator detection in human sera |
| title_fullStr | Development and validation of protein microarray technology for simultaneous inflammatory mediator detection in human sera |
| title_full_unstemmed | Development and validation of protein microarray technology for simultaneous inflammatory mediator detection in human sera |
| title_short | Development and validation of protein microarray technology for simultaneous inflammatory mediator detection in human sera |
| title_sort | development and validation of protein microarray technology for simultaneous inflammatory mediator detection in human sera |
| url | https://eprints.nottingham.ac.uk/40717/ https://eprints.nottingham.ac.uk/40717/ https://eprints.nottingham.ac.uk/40717/ |